Bse3D I

Enzyme nameBse3D I
PrototypeBsrDI
Known analogsBseMI, BsrDI
SKUSE-E253
Turbo versionNot available
High-concentration versionNot available
Restriction typeType IISiType IIS enzymes recognize asymmetric DNA sequences and cleave outside the recognition site at a defined distance. They are widely used in Golden Gate assembly.
Recognition site
5'… GCAATGNN▼ …3'
3'… CGTTAC▲NN …5'
SourceBacillus stearothermophilus 3D
Optimal bufferSE-buffer G (10 mM Tris-HCl (pH 7.6 at 25°C); 10 mM MgCl2; 50 mM NaCl; 1 mM DTT.)
Optimal temperature60 °C
Inactivation temperature80 °C
Buffer activity
BGOWYROSE
10100255075100
Unit definitionOne unit of the enzyme is the amount required to hydrolyze 1 μg of Lambda DNA in 1 hour at 60°C in a total reaction volume of 50 μl.
Assayed onLambda DNA
Storage conditions10 mM Tris-HCl (pH 7.5); 200 mM NaCl; 0.1 mM EDTA; 7 mM 2-mercaptoethanol; 50% glycerol. Store at -20°C.
LigationAfter 5-fold overdigestion with enzyme more than 90% of the DNA fragments can be ligated and recut.
Nonspecific hydrolysisNo nonspecific activity was detected after incubation of 1 μg of Lambda DNA with 10 u.a. of enzyme for 16 hours at 60°C.
Methylation sensitivitynot tested
Reagents supplied with enzyme10 X SE-buffer G
References
  3. Unpublished data Abdurashitov, M.A., Bondar, T.S., Shevchenko, A.V., Dedkov, V.S., Degtyarev, S.K Unpublished observations (1996).
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